You are planning an experimental strategy for isolating your favorite gene and c

Question

You are planning an experimental strategy for isolating your favorite gene and cloning it int o a bacterial vector. You decide to use the E. coli strain MM294 as your plasmid host. You begin the experiment by streaking the stock culture on LB/amp and LB/kan plates as well as an LB plate. When you check your plates the next morning, the LB plate has confluent (strong) growth, the LB/amp plate has approximately 50 colonies and the LB/kan plate has no growth. Are these the results that you anticipated? How do these results influence your experiment?

Explanation / Answer

there are few doubts in the question. did you isolate the gene and clone it into the vector and transform the E.coli MM294 strain culture?

However, if you have done that and then you are using the stock culture to plate on various media the results seen by you can conclude the following results:

when plated only on LB plates the confluent growth indicates that all bacterial cells grew on plate with as well as without the plasmid.

on LB+ amp= 50 colonies indicates that ampicilling resistant bacterial cells have grown. (the plasmid vector contains amp resistance marker thus, the colonies containing the plasmid and hence the favorite cloned gene will grow on the plate).

onLB+Kan= no growth indicates that the plasmid doesnt contain kanamycin resistance marker and thus, there is no growth.

we can pick up the colony from LB+amp and further purify the strain and test for the presence of the cloned gene.

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